Graduation Year

2008

Document Type

Dissertation

Degree

Ph.D.

Degree Granting Department

Molecular Pharmacology and Physiology

Major Professor

Marcia N. Gordon, Ph.D.

Keywords

Transgenic mouse model, Cell therapy, Abeta, Neprilysin, Protease

Abstract

The accumulation of amyloid-ß; protein (Aß) in Alzheimer's disease (AD) is a well known pathological event. Decreasing the production or increasing the degradation of Aß; is therefore thought to serve as a potential therapeutic intervention in AD. Recent in vitro and in vivo studies have suggested that certain proteases may be involved in the catabolism of Aß; and defects in the degradation of Aß; could contribute to AD disease progression. Studies implicating the homing of monocytes to regions of CNS damage have led to the idea that it may be possible to use genetically modified monocytes to carry exogenous genes of interest into the brain or other organs for the purposes of gene therapy.

To determine the time course of monocyte recruitment into the brain during the neurodegenerative damage characteristic of Alzheimer's disease, we used transplanted GFP labeled bone marrow monocytes to characterize the kinetics that peripheral monocytes display once injected into the circulation. We determined the half life of bone marrow derived monocytes after one injection into the peripheral circulation, and found this time to be 1.5 hours post injection. We also examined the effects of the APP+PS1 transgene on the recruitment of peripheral monocytes and showed that these cells are actively recruited to the brains in AD transgenic mouse models compared to non transgenic mice. As an approach to increase expression of NEP in a transgenic mouse model of AD, we developed an ex vivo gene therapy method utilizing bone marrow monocytes from GFP mice.

These monocytes were transfected with a NEP construct designed to express either a secreted form of NEP or a form which lacks any enzyme activity. Monocytes were administered through a microvascular port twice a week for two months and we observed recruitment of bone marrow-derived monocytes into the CNS. In addition, we found significant reductions in both Aß and Congo red staining in the NEP-S injected mice only. These studies show that putting monocytes together with an amyloid degrading enzyme such as neprilysin offers a powerful novel therapeutic tool for the treatment of AD.

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